LIFESCI 23L Study Guide - Final Guide: Polyacrylamide Gel Electrophoresis, Sanger Sequencing, Haplotype
Document Summary
Dna sequencing ddntp used ddntp - h at 3" position to stop synthesis. Dna template, primer w/ label for initiation, dntps, dna pol. Color tag of fragment indicates which nuc it carries at 3" end. Amplify control region/hypervariable segment i of dna during pcr. 16569 bp total (focus on 600 bp in hv region) 50,000 to 200,000 years ago ancestors migrated from afria. Indivs w/ same haplotype as you and other similar haplotypes. Chelex binds mg2+ to prevent dna degradation by enzymes/dnases. Melt by heating, anneal at lower temp, elongate. 35 cycles - 1 billion copies of region of interest. Forward or reverse primer that performs the sequencing. Use gel electrophoresis (sds page) or gel filtration. Est molecular weight of protein in native state. Observe sympathetic ns activation/ flight or fight response. Correlate fitness level w/ amt of daily exercise. Measures movement of air in and out of lungs. Obtain lung capacities and volumes, normal tidal volume, flow volume loop.
